CN1891831A - Method for preparing rhamnolipid - Google Patents
Method for preparing rhamnolipid Download PDFInfo
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- CN1891831A CN1891831A CN 200510040858 CN200510040858A CN1891831A CN 1891831 A CN1891831 A CN 1891831A CN 200510040858 CN200510040858 CN 200510040858 CN 200510040858 A CN200510040858 A CN 200510040858A CN 1891831 A CN1891831 A CN 1891831A
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Abstract
The invention discloses a manufacture method for rhamnolipid that includes the following steps: filtering fungus, inoculating into culture medium after taking dipping preprocess to the sample getting from oil field sewage or grease, inoculating on blood agar plate for culturing, selecting single colony, inoculating on blue agar plate for culturing, selecting the colony having blue halo to the bevel for microbial conserving; fermenting the primary screening strain in fermenting culture medium, testing the oil drainage activity, emulsibility and fermenting liquid surface tension, selecting target strain; taking ferment to the selected strain, and optimizing the fermenting culture medium, fermenting condition and strain to gain rhamnolipid fermenting liquid; distilling rhanmolipid raw product from the rhamnolipid fermenting liquid. The invention has high yield, good heavy hydrocarbon degradation effect, and improves the selecting accuracy.
Description
One, technical field
The invention discloses a kind of preparation method's of glycolipid type biological surfactant, particularly a kind of rhamnolipid preparation method.
Two, background technology
Rhamnolipid belongs to a kind of glycolipid type biological surfactant, except that having effects such as reducing surface tension, stable emulsion and increase foam, also have general synthetic surfactant not available nontoxic, can biological degradation etc. advantage.Therefore, can be widely used in the every field of industry, agricultural, medicine and daily life, be particularly suitable for petroleum industry and field of environment engineering, as the biological viscosity reduction of oil, improve the biological restoration of oil recovery factor, heavy oil contaminated soil etc.In addition, as natural additive, rhamnolipid also more and more is subjected to people's favor in industrial aspect such as foodstuffs industry, fine chemistry industry, medicine and agriculturals.
In the prior art, the existing report that adopts the synthetic rhamnolipid of microbial method.In the preparation process of rhamnolipid, the screening of bacterial classification is the most important, present widely used method is the blood agar flat band method, it is by the cultivation to generation bacterium in the former state, make the red corpuscle generation haemolysis in the blood agar culture-medium, the size of the haemolysis loop diameter that forms by periphery of bacterial colonies is relatively carried out rapid screening again, obtain to prepare the good quality strain of high yield rhamnolipid, then good quality strain was inserted in the different time and contain in the fermention medium of different carbon sources, with the metabolism of promotion thalline, thereby obtain rhamnolipid.Biotechnol.Prog.2002,18,1277～1281 " RhamnolipidBiosurfactant Production by Strains of Pseudomonas aeruginosa Using Low-CostRaw Materials " disclose a kind of preparation method of rhamnolipid, it is in the soil of gasoline, contaminated by diesel oil, dilution coating process by the blood agar flat board, screening produces the generation bacterium of rhamnolipid, obtaining with crude oil is two strain quality strains of sole carbon source growth, and further analysis draws the high yield bacterium of DS10-129 bacterial strain for the preparation rhamnolipid.Serving as to produce in the process that bacterium produces rhamnolipid with the DS10-129 bacterial strain, in the different time, insert the substratum that contains different carbon sources and carry out fermentation culture, cultivate 288 hours approximately after, can obtain the high yield rhamnolipid of 4.31g/L.The above-mentioned method for preparing rhamnolipid has following deficiency: 1, adopt the dilution coating process of blood agar flat board to produce the rhamnolipid bacterial strain screening, though can see the generation of haemolysis circle intuitively, but the haemolysis circle is irregular, some compositions in the blood or assorted bacterium usually also can produce similar haemolysis circle, what of product tensio-active agent are very difficult size by the haemolysis circle judge in the actually operating, and in culturing process, some compositions in the blood or assorted bacterium usually also can produce similar haemolysis circle, and culture medium preparation and use are caused living contaminants; 2, this screening method workload when the dilution coating is big, the poor growth of bacterial strain when being sole carbon source with crude oil, thereby cause the cycle of screening longer; 3, this method needs to use different carbon sources in the fermentation culture process, promptly needs feed supplement, has therefore increased operation sequence, and the output of this bacterial strain is still low excessively simultaneously, is unfavorable for industrial applications.
Three, summary of the invention
The object of the present invention is to provide that a kind of bacterial screening technology is simple, quality good, fermentation period is short, the preparation method of the low rhamnolipid of productive rate height and production cost.
The technical scheme that realizes the object of the invention is: a kind of preparation method of rhamnolipid may further comprise the steps:
1.1 screening: cultivated 22～24 hours to inserting proliferated culture medium after the sample immersion pre-treatment of taking from oilfield sewage or greasy filth at bacterial classification under 25～35 ℃ the envrionment temperature, adopting method of scoring to be inoculated in through the nutrient solution of multiplication culture cultivated on the blood agar flat board 24～72 hours, single bacterium colony of picking haemolysis circle, and be inoculated in and cultivated on the blue agar plate 24～120 hours, the bacterium colony that blue haloing appears in picking inserts the inclined-plane and does culture presevation; After fermenting 72～120 hours in the primary dcreening operation bacterial strain access fermention medium that is deposited in the slant medium, measure its oil extraction activity, emulsifying property and fermented liquid surface tension, choose the purpose bacterial strain then, and deposit kind;
1.2 the optimization of zymotechnique: will ferment through the bacterial strain that screening is chosen, at first carry out fermention medium optimization during the fermentation, carbon source, nitrogenous source, inorganic salt and each substrates quantity to fermenting process are optimized selection, determine optimum fermention medium by investigating rhamnolipid output, fermented liquid surface tension variations, fermentation period length, fermentation costs and the fermenting process control difficulty of bacterial strain under each condition; Secondly, carry out fermentation condition optimization, to leavening temperature be 3～11 at 20～50 ℃, pH value, the condition of dissolved oxygen, stirring velocity and bio-reactor liquid amount and seed culture, inoculum size 1%～10% is optimized; Carry out bacterial classification optimization at last and obtain rhamnolipid fermentation liquor;
1.3 extraction to rhamnolipid fermentation liquor: with fermented liquid behind the bactofugation body, remove by filter insoluble impurities and residual solution wax, pH value of solution=6.0～6.5, add extraction solvent, stir, extract separatory, merge organic phase, wash with equal-volume, remove solvent under reduced pressure with Rotary Evaporators, obtain the rhamnolipid crude samples.
Among the preparation method of rhamnolipid of the present invention, will be linked into liquefaction glycerine through the bacterial strain that screening is chosen is to carry out fermentation technology optimization in the sole carbon source.
Among the preparation method of rhamnolipid of the present invention, the rhamnolipid crude samples is carried out column chromatography purification: at first adorn post with wet method, with chloroform the rhamnolipid crude samples is dissolved, slowly add the silica gel that is dissolved in the chloroform then, gently vibrate silicagel column while adding, add a cover core after silicagel column installs, and carry out balance with chloroform; Slowly and continuously go up sample then, wash to the colour band appearance, launch, emit liquid, launch fully to begin to be moved down near the column bottom up to yellow colour band from the bottom with chloroform/methanol with chloroform; Use the chloroform wash-out again,, collect the material that is come out by wash-out from the below, boil off solvent from yellow colour band; Repeat above-mentioned column chromatography purification process, obtain purer rhamnolipid sample.
Among the preparation method of rhamnolipid of the present invention, purer rhamnolipid sample is further purified: rhamnolipid sample that will be purer dissolves with less water, extracts with chloroform/methanol, after removing solvent under reduced pressure, with acetone product is dissolved again, concentrating under reduced pressure carries out recrystallization.
Compared with prior art, the present invention has following remarkable advantage: (1) the present invention screens the ability that strain Pseudomonas sp.L-21 has strong degrading crude oil, degradation rate to crude oil can reach 59%, and degraded back crude oil is by good dispersion, emulsification or become the water-insoluble sedimentation; (2) the present invention adopts blood agar-blue agar plate screening method, overcomes the shortcoming that in the past only relied on the bio-surfactant hemolytic to judge, has improved the screening accuracy, has saved screening time; (3) output height of the present invention, production peak can reach 23g/L, and 25 ℃ can make the fermented liquid surface tension be reduced to 25.7mN/m; (4) the heavy hydrocarbon degradation effect of the bacterial strain that screens is fairly obvious, and the product rhamnolipid has significant emulsifying effect, thereby the environment remediation effect that is used for pollution by oil soil is obvious.
Four, description of drawings
Fig. 1 is the preparation method's of a rhamnolipid of the present invention schema.
Fig. 2 is the schema of preparation method's bacterial screening of rhamnolipid of the present invention.
Five, embodiment
Below in conjunction with accompanying drawing the present invention is described in further detail.
Embodiment 1: in conjunction with the accompanying drawings, the preparation method of rhamnolipid of the present invention may further comprise the steps:
1.1 bacterial screening: to inserting proliferated culture medium after the sample immersion pre-treatment of taking from oilfield sewage or greasy filth, adopting method of scoring to be inoculated on the blood agar flat board through the nutrient solution of multiplication culture cultivates, single bacterium colony of picking haemolysis circle, and be inoculated on the blue agar plate and cultivate, the bacterium colony that blue haloing appears in picking inserts the inclined-plane and does culture presevation; The primary dcreening operation bacterial strain that is deposited in the slant medium is inserted fermentation back mensuration its oil extraction activity, emulsifying property and fermented liquid surface tension in the fermention medium, choose the purpose bacterial strain then, and deposit kind; Promptly from oilfield sewage or greasy filth, choose sample, after sterilized water immersion pre-treatment in 2 hours, after the proliferated culture medium of liquid access is just soaked in taking-up, it is streak culture in dull and stereotyped 30 hours of blood agar after 30 ℃ of temperature environments are cultivated 24 hours, will single bacterium colony of haemolysis circle appears, the single bacterium colony that occurs is connected to blue agar plate again cultivated 48 hours, occur then being preserved on the slant medium 30 ℃ of temperature environments cultivation preservations 12 hours than the bacterium colony of Da Lan circle 30 ℃ of temperature environments.Insert then cultivate in the seed culture medium after, 5% is inoculated in fermention medium carries out 30 ℃ of temperature environment fermentation culture 48 hours, carry out oil extraction active testing, emulsifying property test and capillary mensuration then, obtain oil extraction circle maximum, the strain Pseudomonas sp.L-21 that surface tension is minimum.
Substratum wherein is as follows: bacterium, actinomycetes enrichment medium (Bacteria and ActinomyceteEnriched Media): cetyl trimethylammonium bromide 2g, (NH
4)
2SO
42g, Na
2HPO
42H
2O 2g, KH
2PO
42g, MgSO
47H
2O 0.5g, CaCl
22H
2O 0.005g liquefied petrolatum 20mL, deionized water is settled to 1L, pH=7～7.5.121 ℃, behind the sterilization 20min, be cooled to about 50 ℃, add the aseptic nystatin 0.05g/L aqueous solution.
Mould and yeast enrichment medium (Mold and Yeast Enrichment Media): cetyl trimethylammonium bromide 2g, (NH
4)
2SO
42g, Na
2HPO
42g, KH
2PO
43g, MgSO
47H
2O 0.5g, CaCl
22H
2O 0.005g, yeast powder 0.1g, liquefied petrolatum 20mL, deionized water is settled to 1L, pH=5.1～5.4.121 ℃ of sterilization 20min are cooled to about 50 ℃, add the aseptic tsiklomitsin 0.035g/L aqueous solution.
Blood agar culture-medium: defiber sheep blood 50mL, extractum carnis 0.3g, peptone 1g, yeast powder 0.01g, fixed molten with deionized water to 1L.At first take by weighing other composition except that sheep blood, heating makes and is dissolved in the 950mL water, 121 ℃ of sterilization 20min, take out, wait to be chilled to about 60 ℃, add the defiber sheep blood 50mL behind filtering with microporous membrane, pour plate is stand-by, and toppling process will prevent that bubble from producing and agar solidifies.
Blue nutrient agar: cetyl trimethylammonium bromide 5g, methylene blue 0.02g, extractum carnis 0.3g, peptone 1g, yeast powder 0.01g, fixed molten with deionized water to 1L.At first take by weighing cetyl trimethylammonium bromide and methylene blue other composition in addition, heating makes and is dissolved in the deionized water.Take by weighing the 5g cetyl trimethylammonium bromide then, the 0.02g methylene blue is dissolved in the less water.Treat that the former is cooled to about 60 ℃～70 ℃, add the latter, supply deionized water, slowly stir, need not sterilize, culture dish is cleaned and oven dry, directly pour plate is stand-by.
1.2 the optimization of zymotechnique: it is that sole carbon source ferments that the bacterial strain that screening is obtained is linked into liquefaction glycerine, obtains rhamnolipid fermentation liquor, and promptly fermentation technology optimization comprises that bacterial classification optimization, fermention medium optimization, fermentation condition optimization three parts carry out.At first carry out fermention medium optimization, carbon source, nitrogenous source, inorganic salt and each substrates quantity to fermenting process are optimized selection, determine optimum fermention medium by investigating rhamnolipid output, fermented liquid surface tension variations, fermentation period length, fermentation costs and the fermenting process control difficulty of bacterial strain under each condition; Secondly, carry out fermentation condition optimization, the condition of leavening temperature in 20～50 ℃, pH value 3～11, dissolved oxygen, stirring velocity and bio-reactor liquid amount and seed culture, inoculum size 1%～10% is optimized; Carry out bacterial classification optimization at last, method for mutation breeding improvement bacterial classification is adopted in bacterial classification optimization, makes output, strain growth and metabolism state be fit to the suitability for industrialized production needs.
Pseudomonas sp.L-21 is inoculated in the seed culture medium seed culture medium: liquefied petrolatum 20mL, (NH
4)
2SO
42g, Na
2HPO
42H
2O 2g, KH
2PO
42g, MgSO
47H
2O 0.5g, CaCl
22H
2O 0.005g, extractum carnis 1g is settled to 1L with deionized water, and transferring pH is 7.0～7.2,121 ℃ of autoclaving 20min.30 ℃, 200r/min shaking table shaking culture 24h.Then seed culture medium is inserted fermention medium, 30 ℃, 200r/min cultured continuously 84h with 5% inoculum size.
Fermention medium: glycerine 30mL, (NH
4)
2SO
42g, Na
2HPO
42H
2O 3g, KH
2PO
41g, MgSO
47H
2O 0.5g, CaCl
22H
2O 0.005g is settled to 1L with deionized water, and transferring pH is 8.Culture temperature: 30 ℃, shaking speed: 200r/min; Shake bottled liquid measure: 30～100mL; Inoculum size: 4%; Incubation time: 72～80h; The seed culture time: 21h.
1.3 obtain rhamnolipid fermentation liquor through bacterial screening and fermentation optimization, now rhamnolipid fermentation liquor is extracted, can judge tentatively that from the bacterial screening process Pseudomonas sp.L-21 can secrete the outer glycolipid type biological surfactant of born of the same parents during the fermentation.Therefore with the 1L fermented liquid after the centrifugal 30min of 4000r/min removes thalline, remove by filter insoluble impurities and residual solution wax.Filtrate is used 10%H
2SO
4Solution is transferred pH value of solution=6.0～6.5, adds the chloroform/methanol (V/V=2: 1) mixed solvent, magnetic agitation 1h of 2L volume at twice, separating funnel extraction separatory merges organic phase, washes with equal-volume, under 45 ℃, remove solvent under reduced pressure, obtain the rhamnolipid crude samples with Rotary Evaporators.
The rhamnolipid crude samples is carried out column chromatography purification: at first be the dress post, adopt wet method dress post, thick product is dissolved with chloroform.At first pack in the post chloroform of 2/3 volume slowly adds the silica gel that is dissolved in the chloroform then, gently vibrates silicagel column while adding.Simultaneously, keep solvent to flow out from the below with the speed of 1/s.Note to have the globule to sneak in dress post process silicagel column, silica gel and the solvent.Add a cover core after silicagel column installs, and carry out balance with chloroform.Go up sample and wash-out then, crude product A is dissolved in about 10mL chloroform, crossing length-to-diameter ratio as stated above is 60cm * 3cm silicagel column, it is slow and continuously that last sample process is wanted, can not the disturbance liquid level, otherwise will influence the colour band edge, be unfavorable for the carrying out of wash-out and purge process, even cause the chromatography failure.Wash to the colour band appearance with chloroform earlier, use chloroform/methanol (V/V=8: 1) launch, emit liquid from the bottom, launch fully to begin to be moved down into then near the column bottom up to yellow colour band with 1/min; Begin to use the chloroform wash-out, begin to come out to begin to collect from the below by wash-out from yellow colour band, every 10mL collects once, makes elution curve.Collection also boils off solvent, obtains purer rhamnolipid product.Repeat said process, can obtain purer rhamnolipid product sample.
The present invention can also be further purified: will dissolve with less water through the rhamnolipid sample of column chromatography purification, with the chloroform/methanol of about 2 times of volumes (V/V=8: 1) extraction, remove solvent under reduced pressure after, with acetone that product is molten down, concentrating under reduced pressure is placed and is made the product recrystallization, gets more pure products.
The glycolipid sample that purification obtains is analyzed through ESI-MS and is determined, has multiple different structure rhamnolipid component in the extract, mainly by Rh
2C
10C
10, RhC
10C
10Two kinds of materials constitute, and account for about 72.3% of total amount greatly, and all the other various rhamnolipid components account for about 27.7%; In isomers, for the rhamnolipid of the beta-hydroxycarboxylic acids that contains two different carbon chain lengths, glycosyl and short chain beta-hydroxycarboxylic acids bonded rhamnolipid component concentration are higher than long-chain and rhamanopyranosyl bonded rhamnolipid component.In two rhamnolipids (dirhamnolipid) structure, away from the pyranoid ring texture ratio of lipid acid long-chain near the lipid acid long-chain or to be arranged in single rhamnolipid (monorhamnolipid) structure rhamanopyranosyl stable.Two rhamnolipid content are more than single rhamnolipid in two kinds of main rhamnolipid components.Rh wherein
2C
10C
10(MW:650) the rhamnolipid component concentration accounts for 42.36% of total abundance, and structure is RhC
10C
10(MW:504) rhamnolipid component concentration accounts for 29.84% of total abundance.
Embodiment 2: the preparation method of rhamnolipid of the present invention, from the greasy dirt of oil field, sample, obtain bio-surfactant and produce bacterium by the dull and stereotyped composite screen lectotype of enrichment culture, blood agar (primary dcreening operation) and blue gel (multiple sieve) screening, and through the ultraviolet mutagenesis breeding improvement, the excellent species that obtains is preserved in the PDA inclined-plane.By fermentation culture, i.e. fermention medium: whiteruss or glucose or soya-bean oil or diesel oil or glycerine 15-35mL, (NH
4)
2SO
42-2.5g, Na2HPO
42H
2O3-3.8g, KH
2PO
41g, MgSO
47H
2O 0.5g, CaCl
22H
2O 0.005g is settled to 1L with deionized water, and accent pH is 7.5-8.Culture condition: temperature: 26～36 ℃, shaking speed: 14～240r/min; Erlenmeyer flask liquid amount: 50-100mL; The seed culture time: 21h; Inoculum size: 5～10%; Incubation time: 48～158h, fermented liquid be with chloroform and methanol extraction, obtains head product mixing rhamnolipid after boiling off solvent.
After silicagel column separates, obtain a kind of yellow needle-like crystal, in conjunction with TLC, ESI-MS, FI-IR,
1H NMR,
13Technology such as C NMR, GC-MS are identified crystal.It is mainly by Rh
2C
10C
10, RhC10C
10Two kinds of materials constitute, and account for greatly about the 72.1-72.3% of rhamnolipid amount of the mixture, and all the other various rhamnolipid components account for about 27.9-27.7%; Rh wherein
2C
10C
10Rhamnolipid component abundance is 42.36-42.41%, and structure is RhC
10C
10Rhamnolipid component abundance be 29.84-30.15%.
Claims (4)
1, a kind of preparation method of rhamnolipid is characterized in that may further comprise the steps:
1.1 screening: cultivated 22～24 hours to inserting proliferated culture medium after the sample immersion pre-treatment of taking from oilfield sewage or greasy filth at bacterial classification under 25～35 ℃ the envrionment temperature, adopting method of scoring to be inoculated in through the nutrient solution of multiplication culture cultivated on the blood agar flat board 24～72 hours, single bacterium colony of picking haemolysis circle, and be inoculated in and cultivated on the blue agar plate 24～120 hours, the bacterium colony that blue haloing appears in picking inserts the inclined-plane and does culture presevation; After fermenting 72～120 hours in the primary dcreening operation bacterial strain access fermention medium that is deposited in the slant medium, measure its oil extraction activity, emulsifying property and fermented liquid surface tension, choose the purpose bacterial strain then, and deposit kind;
1.2 the optimization of zymotechnique: will ferment through the bacterial strain that screening is chosen, at first carry out fermention medium optimization during the fermentation, carbon source, nitrogenous source, inorganic salt and each substrates quantity to fermenting process are optimized selection, determine optimum fermention medium by investigating rhamnolipid output, fermented liquid surface tension variations, fermentation period length, fermentation costs and the fermenting process control difficulty of bacterial strain under each condition; Secondly, carry out fermentation condition optimization, the condition of leavening temperature in 20～50 ℃, pH=3～11, dissolved oxygen, stirring velocity and bio-reactor liquid amount and seed culture, inoculum size 1%～10% is optimized; Carry out bacterial classification optimization at last and obtain rhamnolipid fermentation liquor;
1.3 extraction to rhamnolipid fermentation liquor: with fermented liquid behind the bactofugation body, remove by filter insoluble impurities and residual solution wax, pH value of solution=6.0～6.5, add extraction solvent, stir, extract separatory, merge organic phase, wash with equal-volume, remove solvent under reduced pressure with Rotary Evaporators, obtain the rhamnolipid crude samples.
2, the preparation method of rhamnolipid according to claim 1 is characterized in that it is to carry out fermentation technology optimization in the sole carbon source that the bacterial strain that will choose through screening is linked into liquefaction glycerine.
3, the preparation method of rhamnolipid according to claim 1 and 2, it is characterized in that the rhamnolipid crude samples is carried out column chromatography purification: at first adorn post with wet method, with chloroform the rhamnolipid crude samples is dissolved, slowly add the silica gel that is dissolved in the chloroform then, gently vibrate silicagel column while adding, add a cover core after silicagel column installs, and carry out balance with chloroform; Slowly and continuously go up sample then, wash to the colour band appearance, launch, emit liquid, launch fully to begin to be moved down near the column bottom up to yellow colour band from the bottom with chloroform/methanol with chloroform; Use the chloroform wash-out again,, collect the material that is come out by wash-out from the below, boil off solvent from yellow colour band; Repeat above-mentioned column chromatography purification process, obtain purer rhamnolipid sample.
4, the preparation method of rhamnolipid according to claim 3, it is characterized in that purer rhamnolipid sample is further purified: rhamnolipid sample that will be purer dissolves with less water, extract with chloroform/methanol, after removing solvent under reduced pressure, with acetone product is dissolved again, concentrating under reduced pressure carries out recrystallization.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182560B (en) * | 2007-11-29 | 2011-04-27 | 湖南大学 | Method for enhancing yield of rhamnolipid produced by copper green pseudomonas |
| CN101177696B (en) * | 2007-11-05 | 2011-06-08 | 大庆沃太斯化工有限公司 | Industrial preparation method of rhamnolipid biological fermentation liquor |
| CN102174609A (en) * | 2011-01-21 | 2011-09-07 | 天津工业生物技术研究所 | Field fermentation system for oil displacement biological surfactant |
| CN102766172A (en) * | 2011-09-19 | 2012-11-07 | 大庆沃太斯化工有限公司 | Industrial production method of rhamnolipid biosurfactant dry powder |
| CN103202295A (en) * | 2010-03-30 | 2013-07-17 | 湖州紫金生物科技有限公司 | Application of rhamnolipid as biological pesticide and biological insecticide |
| CN105670969A (en) * | 2016-03-10 | 2016-06-15 | 中国科学院水生生物研究所 | Method and application for screening bacteria capable of relieving constructed wetland blocking |
| US9884883B2 (en) | 2015-01-12 | 2018-02-06 | Logos Technologies, Llc | Production of rhamnolipid compositions |
| CN108191930A (en) * | 2018-01-22 | 2018-06-22 | 中国科学院沈阳应用生态研究所 | A kind of method of rhamnolipid product in extraction zymotic fluid |
| CN110819570A (en) * | 2019-11-27 | 2020-02-21 | 中秀科技股份有限公司 | Blood agar plate and preparation method thereof |
| CN111892254A (en) * | 2020-09-02 | 2020-11-06 | 浙江一清环保工程有限公司 | Method for producing rhamnolipid by fermenting kitchen wastewater and fish meal wastewater in resource utilization mode |
| US10829507B2 (en) | 2017-02-06 | 2020-11-10 | Stepan Company | Decolorization of concentrated rhamnolipid composition |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101177696B (en) * | 2007-11-05 | 2011-06-08 | 大庆沃太斯化工有限公司 | Industrial preparation method of rhamnolipid biological fermentation liquor |
| CN101182560B (en) * | 2007-11-29 | 2011-04-27 | 湖南大学 | Method for enhancing yield of rhamnolipid produced by copper green pseudomonas |
| CN103202295B (en) * | 2010-03-30 | 2015-01-07 | 湖州紫金生物科技有限公司 | Application of rhamnolipid as biological pesticide and biological insecticide |
| CN103202295A (en) * | 2010-03-30 | 2013-07-17 | 湖州紫金生物科技有限公司 | Application of rhamnolipid as biological pesticide and biological insecticide |
| CN102174609A (en) * | 2011-01-21 | 2011-09-07 | 天津工业生物技术研究所 | Field fermentation system for oil displacement biological surfactant |
| CN102174609B (en) * | 2011-01-21 | 2013-04-10 | 天津工业生物技术研究所 | Field fermentation system for oil displacement biological surfactant |
| CN102766172A (en) * | 2011-09-19 | 2012-11-07 | 大庆沃太斯化工有限公司 | Industrial production method of rhamnolipid biosurfactant dry powder |
| US9884883B2 (en) | 2015-01-12 | 2018-02-06 | Logos Technologies, Llc | Production of rhamnolipid compositions |
| CN105670969A (en) * | 2016-03-10 | 2016-06-15 | 中国科学院水生生物研究所 | Method and application for screening bacteria capable of relieving constructed wetland blocking |
| US10829507B2 (en) | 2017-02-06 | 2020-11-10 | Stepan Company | Decolorization of concentrated rhamnolipid composition |
| CN108191930A (en) * | 2018-01-22 | 2018-06-22 | 中国科学院沈阳应用生态研究所 | A kind of method of rhamnolipid product in extraction zymotic fluid |
| CN108191930B (en) * | 2018-01-22 | 2021-02-09 | 中国科学院沈阳应用生态研究所 | Method for extracting rhamnolipid product from fermentation liquor |
| CN110819570A (en) * | 2019-11-27 | 2020-02-21 | 中秀科技股份有限公司 | Blood agar plate and preparation method thereof |
| CN111892254A (en) * | 2020-09-02 | 2020-11-06 | 浙江一清环保工程有限公司 | Method for producing rhamnolipid by fermenting kitchen wastewater and fish meal wastewater in resource utilization mode |
| CN111892254B (en) * | 2020-09-02 | 2022-08-16 | 浙江一清环保工程有限公司 | Method for producing rhamnolipid by fermenting kitchen wastewater and fish meal wastewater in resource utilization mode |
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